Normalization techniques for high‐throughput screening by infrared matrix‐assisted laser desorption electrospray ionization mass spectrometry

Mass spectrometry (MS) is an effective analytical tool for high‐throughput screening (HTS) in the drug discovery field. Infrared matrix‐assisted laser desorption electrospray ionization (IR‐MALDESI) MS is a high‐throughput platform that has achieved analysis times of sub‐seconds‐per‐sample. Due to the high‐throughput analysis speed, methods are needed to increase the analyte signal while decreasing the variability in IR‐MALDESI‐MS analyses to improve data quality and reduce false‐positive hits. The Z‐factor is used as a statistic of assay quality that can be improved by reducing the variation of target ion abundances or increasing signal. Herein we report optimal solvent compositions for increasing measured analyte abundances with direct analysis by IR‐MALDESI‐MS. We also evaluate normalization strategies, such as adding a normalization standard that is similar or dissimilar in structure to the model target drug, to reduce the variability of measured analyte abundances with direct analyses by IR‐MALDESI‐MS in both positive and negative ionization modes.